CN105223366B - Micropipe for blood grouping, one-piece ABO/RhD blood typing test card for blood grouping and preparation thereof - Google Patents
Micropipe for blood grouping, one-piece ABO/RhD blood typing test card for blood grouping and preparation thereof Download PDFInfo
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- CN105223366B CN105223366B CN201510601037.4A CN201510601037A CN105223366B CN 105223366 B CN105223366 B CN 105223366B CN 201510601037 A CN201510601037 A CN 201510601037A CN 105223366 B CN105223366 B CN 105223366B
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/80—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving blood groups or blood types or red blood cells
Abstract
The invention discloses a micropipe for blood grouping, a one-piece ABO/RhD blood typing test card for blood grouping and a preparation thereof. The micropipe for blood grouping includes a gel, an antibody working solution and a serum separation gel from bottom to top in order. The antibody working solution comprises a blood group antibody and an antibody diluent which can make the blood group antibody maintain activity, and the serum separation gel having a specific gravity of 1.075-1.077 can perform thixotropy to turn the solid into the fluid in the presence of centrifugal force of 150 G to 300 G. The specific gravity of the antibody diluents is more than that of the serum separation gel and less than that of the erythrocyte. The one-piece ABO/RhD blood typing test card prepared by the micropipe can add peripheral blood directly to test, and has no need to test the blood sample after diluting the peripheral blood removed the erythrocyte by centrifuging. So that the sensitivity and specificity of the blood type test card are guaranteed, the operation steps of the traditional ABO/RhD blood typing test card are reduced, and the efficiency is improved.
Description
Technical field
The present invention relates to Blood grouping, more particularly to a kind of micro-pipe for Blood grouping, integral type abo/rhd blood group
Sizing detection card and preparation method.
Background technology
Since self-discovery abo blood group, abo blood group is widely used in clinic, and the accurate sizing of abo blood group is extremely important,
It is basis and the guarantee of safe transfusion.Rh blood group system is the important blood group system being only second to abo blood group system, is also red blood cell
A most complicated system in blood group system, the antigen in rh blood group system includes d, c, e etc., wherein d antigen
By force.It is that rhd is positive that erythrocyte surface contains d antigen, and not containing d antigen is that rhd is negative, and rhd negative patient accepts rhd positive blood
After liquid, at least 20% patient can produce immune response to d antigen, haemolysis can be occurred anti-after contacting this antigen in vivo at second
Should, therefore rhd is carried out shaping also particularly significant.
Current abo/rhd blood typing detects that the red blood cell that calorie requirement adds after dilution just can be detected, operation step
Suddenly loaded down with trivial details, inefficiency.
Content of the invention
In order to make up above-mentioned the deficiencies in the prior art, the present invention proposes a kind of micro-pipe for Blood grouping, integral type
Abo/rhd blood typing detection card and preparation method.
The technical problem of the present invention is solved by following technical scheme:
The present invention provide a kind of micro-pipe for Blood grouping be: described micro-pipe from bottom up comprise successively gel,
Antibody working solution and serum separation gel；Containing blood group antibody with make described blood group antibody keep activity in described antibody working solution
Antibody diluent；The proportion of described serum separation gel between 1.075～1.077, under the centrifugal action of 150g～300g
Thixotroping can occur, fluid is become by solid；The proportion of described antibody diluent is more than serum separation gel and is less than red blood cell.
The integral type abo/rhd blood typing detection card that the present invention provides is: has 8 micro-pipes, institute on described detection card
Stating 8 micro-pipes is respectively:
First micro-pipe: the blood group antibody for containing in described micro-pipe, and described antibody working solution is anti-a monoclonal antibody；
Second micro-pipe: the blood group antibody for containing in described micro-pipe, and described antibody working solution is anti-b monoclonal antibody；
3rd micro-pipe: the blood group antibody for containing in described micro-pipe, and described antibody working solution is anti-d monoclonal antibody；
4th micro-pipe: up comprise gel, antibody diluent and serum separation gel successively from the bottom of described 4th micro-pipe；
5th micro-pipe～the 8th micro-pipe: repeat the first micro-pipe～the 4th micro-pipe respectively.
The preparation method of the integral type abo/rhd blood typing detection card that the present invention provides, comprises the steps of:
(1) prepare antibody diluent, the proportion of described antibody diluent is more than serum separation gel and is less than red blood cell；
(2) gel swelling: using physiological saline or described antibody diluent, gel is carried out swelling；
(3) prepare antibody working solution: respectively anti-a monoclonal antibody, anti-b monoclonal antibody and anti-d monoclonal antibody are pressed
The antibody of every 1 parts by volume adds the described antibody diluent proportions of 7 parts by volume to become described antibody working solution；
(4) prepare specific gel: add antibody working solution described in 1.2-1.7ml according to swelling gel described in every 1g
Ratio be configured to the anti-monoclonal anti-a gel with igm property, the anti-monoclonal anti-b gel with igm property and tool respectively
There is the anti-monoclonal anti-d gel of igm property；And preparation neutral gum: add 1.2- according to swelling gel described in every 1g
The proportions of antibody diluent described in 1.7ml become neutral gum；Wherein: anti-a antibody titer >=128, anti-b monoclonal resists
Body potency >=128, anti-d antibody titer >=128；
(5) dispense: according to the filling amount of every pipe 45-55 μ l, anti-for described anti-monoclonal a gel, the anti-b of anti-monoclonal are coagulated
Glue, anti-monoclonal anti-d gel, neutral gum are added separately in the first micro-pipe, the second micro-pipe, the 3rd micro-pipe and the 4th micro-pipe, the
Five micro-pipes～the 8th micro-pipe repeats the first micro-pipe～the 4th micro-pipe respectively；
(6) prepare serum separation gel, the proportion of described serum separation gel between 1.075～1.077,150g～
Under the centrifugal action of 2000g, thixotroping can occur, fluid is become by solid；
(7) respectively in the first micro-pipe～the 8th micro-pipe add 0.8g-2g described serum separation gel, 150g-300g from
Heart certain time, described serum separation gel is made to occur thixotroping to be become the interface catching after fluid in described antibody working solution by solid
On；
(8) sealing, labeling and encapsulation.
The beneficial effect that the present invention is compared with the prior art is: the integral type abo/rhd blood typing detection card of the present invention
Periphery whole blood can directly be added to be detected, take out after red blood cell is diluted again without periphery whole blood is carried out with centrifugation
Detected, the sensitivity ensureing blood type test card and specific under the premise of, it is possible to reduce traditional abo/rhd blood group is fixed
The operating procedure of type detection card, improves efficiency.Specifically, it is thin less than red that the serum separation gel proportion that the present invention adopts is more than serum
Born of the same parents, proportion, between 1.075-1.077, can occur thixotroping to become fluid, blood group by solid under the conditions of certain centrifugal force
The good thixotropy of separation gel is to extend the necessary condition that red blood cell and blood group antibody occur agglutinating reaction, the touching of serum separation gel
Denaturation will meet the ample time that red blood cell rests in antibody working solution, in order to avoid red blood cell just can not be reacted with blood group antibody
Through being sunken to ttom of pipe, cause false-negative testing result.
Brief description
Fig. 1 is the schematic diagram of the micro-pipe for Blood grouping in the specific embodiment of the invention.
Specific embodiment
Below against accompanying drawing and with reference to preferred embodiment the invention will be further described.
The present invention provides a kind of micro-pipe for Blood grouping, in a specific embodiment, as shown in figure 1, micro-pipe 1 is the bottom of from
Portion up comprises gel 11, antibody working solution 12 and serum separation gel 13 successively, contains blood group antibody and make in antibody working solution
Blood group antibody keeps the antibody diluent of activity；The proportion of serum separation gel 13 between 1.075～1.077,150g～
Under the centrifugal action of 2000g, thixotroping can occur, fluid is become by solid；The proportion of antibody diluent is more than serum separation gel
And it is less than red blood cell.
The proportion relation of each material is as follows: serum ＜ serum separation gel ＜ antibody specific gravity liquid ＜ red blood cell, preferably real
Apply in example:
, between 1.078-1.082, ph value is between 6.6-6.8 for the proportion of antibody diluent.
Serum separation gel includes each component of following weight portion: cyclopentadiene oligomers 35-45 part, organic gelling agent 12-
25 parts, organic gelling agent dispersant 15-30 part, viscosity reductant 8-12 part and proportion conditioning agent 8-12 part.In preferred embodiment
In, serum separation gel includes each component of following weight portion: 40 parts of cyclopentadiene oligomers, 20 parts of organic gelling agent, organic gel
20 parts of solidifying agent dispersant, 10 parts of viscosity reductant and 10 parts of proportion conditioning agent.
Gel is sephadex, and particle diameter is 40-60 μm.
It is 200g that serum separation gel occurs the centrifugal force of thixotroping, and centrifugation time is 2min.
Under same time (this experiment adopt 2 minutes), design centrifugal force gradient 80g, 100g, 120g, 150g, 180g,
200g, 250g, 300g, 400g, it is demonstrated experimentally that under the centrifugal force more than 150g, serum separation gel can be well red blood cell
Separate with serum, poor less than then effect；And if centrifugal force is higher than 300g although serum separation gel can be by red blood cell and serum
Separate, but because centrifugal force is excessive, red blood cell does not have time enough just to react with the blood group antibody in antibody working solution
Through sinking to ttom of pipe, it is unfavorable for correctly differentiating blood group.
Carry out three groups of experiments under identical centrifugal force, be 150g, 180g and 200g respectively, one time gradient 30s of design,
1min, 1.5min, 2min, it is demonstrated experimentally that be centrifuged 2 minutes effects in 200g preferable.
Described organic gelling agent is dibenzylidene sorbitol；Described organic gelling agent dispersant is polyoxyethylene-polyoxy
Propylene-based block copolymer, 1-Methyl-2-Pyrrolidone, at least one in dmso, dma and dmf；Described viscosity reductant is adjacent benzene two
At least one in formic acid two (2- ethylhexyl) ester and epoxy resin；Described proportion conditioning agent is the dioxy of 5 μm -10 μm of particle diameter
SiClx, titanium dioxide, the chlorinated paraffin of chlorination degree 40%, the chlorinated paraffin of chlorination degree 70%.
The weight ratio of the volume of described antibody working solution and described gel is between 1:3 to 1:5；Described serum separation gel
Addition be 0.8-2g, more optimizedly add 1g.The present invention also provides a kind of integral type abo/rhd blood typing detection card,
In a specific embodiment, detection card has 8 micro-pipes, 8 micro-pipes are respectively:
First micro-pipe: the blood group antibody for containing in the micro-pipe in above-mentioned embodiment, and antibody working solution is anti-a Dan Ke
Grand antibody；
Second micro-pipe: the blood group antibody for containing in the micro-pipe in above-mentioned embodiment, and described antibody working solution is anti-b
Monoclonal antibody；
3rd micro-pipe: the blood group antibody for containing in the micro-pipe in above-mentioned embodiment, and described antibody working solution is anti-d
Monoclonal antibody；
4th micro-pipe: up comprise gel, antibody diluent and serum separation gel successively from the bottom of the 4th micro-pipe, namely
Blood group antibody is not contained in 4th micro-pipe；
5th micro-pipe～the 8th micro-pipe: repeat the first micro-pipe～the 4th micro-pipe respectively.
The present invention also provides a kind of preparation method of integral type abo/rhd blood typing detection card, in specific embodiment
In, comprise the steps of:
(1) prepare antibody diluent, the proportion of described antibody diluent is more than serum separation gel and is less than red blood cell；
(2) gel swelling: using physiological saline or described antibody diluent, gel is carried out swelling；
(3) prepare antibody working solution: respectively anti-a monoclonal antibody, anti-b monoclonal antibody and anti-d monoclonal antibody are pressed
The antibody of every 1 parts by volume adds the described antibody diluent proportions of 7 parts by volume to become described antibody working solution；
(4) prepare specific gel: add antibody working solution described in 1.2-1.7ml according to swelling gel described in every 1g
Ratio be configured to the anti-monoclonal anti-a gel with igm property, the anti-monoclonal anti-b gel with igm property and tool respectively
There is the anti-monoclonal anti-d gel of igm property；And preparation neutral gum: add 1.2- according to swelling gel described in every 1g
The proportions of antibody diluent described in 1.7ml become neutral gum；Wherein: anti-a antibody titer >=128, anti-b monoclonal resists
Body potency >=128, anti-d antibody titer >=128；
(5) dispense: according to the filling amount of every pipe 45-55 μ l, anti-for described anti-monoclonal a gel, the anti-b of anti-monoclonal are coagulated
Glue, anti-monoclonal anti-d gel, neutral gum are added separately in the first micro-pipe, the second micro-pipe, the 3rd micro-pipe and the 4th micro-pipe, the
Five micro-pipes～the 8th micro-pipe repeats the first micro-pipe～the 4th micro-pipe respectively；
(6) prepare serum separation gel, the proportion of described serum separation gel between 1.075～1.077,150g～
Under the centrifugal action of 2000g, thixotroping can occur, fluid is become by solid；
(7) respectively in the first micro-pipe～the 8th micro-pipe add 0.8g-2g described serum separation gel, 150g-300g from
Heart certain time, described serum separation gel is made to occur thixotroping to be become the interface catching after fluid in described antibody working solution by solid
On；
(8) sealing, labeling and encapsulation.
In a preferred embodiment:
Described step (6) specifically includes following steps:
(61) by 35-45 part cyclopentadiene oligomers heating and melting；
(62) organic gelling agent 12-25 part, organic gelling agent dispersant 15- are added in the cyclopentadiene oligomers melting
30 parts and viscosity reductant 8-12 part, kneaded mixture 20-40 minute；
(63) it is cooled to normal temperature (i.e. 20-30 DEG C), add proportion conditioning agent 8-12 part, in the vacuum of 0.1bar-0.5bar
The lower vacuum kneading mixture 20-40 minute of degree.
The process of the described antibody diluent of preparation in described step (1) is as follows: adds 1.2- in every 1l pure water
1.7g sodium chloride, 12-30g glycine, 0.2-0.3g disodium hydrogen phosphate, 0.46-0.56g sodium dihydrogen phosphate, 10-20g cow's serum
Albumin, 12-25g glycerine, 0.45-0.68g methyl p-hydroxybenzoate and 0.1-0.16g propylparaben.
In a preferred embodiment, the preparation method that this integral type abo/rhd blood typing detection blocks comprises as follows
Step:
(1) prepare antibody diluent, the proportion of described antibody diluent is more than serum separation gel and is less than red blood cell, this example
In, antibody diluent proportion is 1.078-1.082, and ph is 6.6, and formula is as follows: add 1.5g sodium chloride in 1l pure water,
20g glycine, 0.26g disodium hydrogen phosphate, 0.52g sodium dihydrogen phosphate, bovine serum albumin(BSA) 10g, glycerine 20g, para hydroxybenzene first
Sour methyl esters 0.6g and propylparaben 0.13g.
(2) gel swelling, is directly carried out swelling using antibody diluent to gel in this example, specifically include following steps:
(21) 2-10ml antibody diluent is added (to be 1g in this example in every 1g gel (adopting sephadex in this example)
Gel adds 2ml antibody diluent), suspended gel, carry out moist heat sterilization, standby after sterilizing.
(22) gel after step (21) sterilizing is placed certain time (being 6 hours in this example), so that gel is naturally sunk
Fall, removes supernatant, the antibody diluent adding volume ratio 1:1.2 (ratio of the gel of sedimentation and antibody diluent) is resuspended,
So it is repeated twice, remove gel breakage fragment, collected even particle size (in this example, particle diameter is in 40-60 μ m)
Swelling gel.
(3) prepare antibody working solution: respectively by anti-a monoclonal antibody, anti-b monoclonal antibody and anti-d monoclonal antibody (this
Example in, three kinds of antibody are purchased from Merck Millipore Corp.) by every 1 parts by volume antibody addition 7 parts by volume antibody diluent ratio
(in this example, specifically, the volume of each antibody is 5ml to example, for diluting the volume integral of the antibody diluent of each antibody
Wei 35ml) it is configured to antibody working solution.
(4) prepare specific gel: add the ratio of 1.2-1.7ml antibody working solution according to the swelling gel of every 1g
(being the antibody diluent that 1g swell gel adds 1.5ml in this example) is configured to have the anti-a of anti-monoclonal of igm property respectively
Gel, there is the anti-monoclonal anti-b gel of igm property and there is the anti-monoclonal anti-d gel of igm property；And prepare neutral
Glue: the ratio adding 1.2-1.7ml antibody diluent according to the swelling gel of every 1g (is that 1g swell gel adds in this example
The antibody diluent of 1.5ml) it is configured to neutral gum；Wherein: anti-a antibody titer >=128, anti-b antibody titer
>=128, anti-d antibody titer >=128.
(5) dispense: according to the filling amount of every pipe 50 μ l, by anti-for described anti-monoclonal a gel, anti-monoclonal anti-b gel, resist
Monoclonal anti-d gel, neutral gum are added separately in the first micro-pipe, the second micro-pipe, the 3rd micro-pipe and the 4th micro-pipe, the 5th micro-pipe
～the eight micro-pipe repeats the first micro-pipe～the 4th micro-pipe respectively.
(6) prepare serum separation gel, specific as follows:
(61) by 40 parts of cyclopentadiene oligomers at 130 DEG C heating and melting.
(62) add organic gelling agent (being dibenzylidene sorbitol in this example) in the cyclopentadiene oligomers melting
20 parts, 20 parts of organic gelling agent dispersant (being polyox-yethylene-polyoxypropylene block copolymer in this example) is with viscosity reductant (in this example
For phthalic acid two (2- ethylhexyl) ester) 10 parts, kneaded mixture 30 minutes.
(63) it is cooled to normal temperature, add 10 parts of proportion conditioning agent (adopting titanium dioxide in this example), vacuum kneading mixture
30 minutes.
(7) add 1g serum separation gel respectively in the first micro-pipe～the 8th micro-pipe, 200g is centrifuged 3 minutes, so that serum is divided
Occur thixotroping to be become by solid from glue to catch after fluid on the interface of antibody working solution.
(8) sealing, labeling and encapsulation, wherein, sealing refers to: the mouth of pipe of the micro-pipe having dispensed is carried out heating power with aluminium foil
Plastic packaging；Labeling refers to: after the completion of sealing, labeling in each micro-pipe, and form integral type abo/rhd blood typing detection card；
Encapsulation refers to: carries out aseptic plastic packaging and mounted box to integral type abo/rhd blood typing detection card.
Design two groups of experiments individually below, the integral type abo/rhd blood typing detection using above example blocks (below
Abbreviation integrated card) to carry out Blood grouping contrast as follows with the abo blood type test card of Changchun Bo Xun company (hereinafter referred to as conventional card):
First group of experiment:
1st, extract unknown blood group blood donor 10 people, everyone extracts peripheral blood 10ml (sample 1～sample 10).
2nd, take above-mentioned peripheral blood 2.5ml to add in the test tube containing coagulant, 300g is centrifuged 2 minutes, take 16 μ l hematocrits red thin
Born of the same parents, add 1984 μ l physiological saline, are configured to 0.8% red cell suspension.
3rd, integrated card and each 50 of conventional card are taken, 200g is centrifuged 2 minutes, tears aluminium film.
4th, it is directly added into 2 μ l peripheral bloods toward in integrated card, add the red cell suspension of 50 μ l simultaneously toward in conventional card.
5th, the conventional card having added red cell suspension is put into 70g in centrifuge to be centrifuged 1 minute, then 100g centrifugation 4 minutes.
6th, the integrated card having added peripheral blood is put into 200g in centrifuge to be centrifuged 2 minutes, then 100g centrifugation 4 minutes.
7th, card taking result of determination, result such as following table one.
The unknown blood group pattern detection result of table one
| Sample | 1 | 2 | 3 | 4 | 5 | 6 | 7 | 8 | 9 | 10 |
| Integrated card | a | o | b | a | b | b | ab | b | o | a |
| Conventional card | a | o | b | a | b | b | ab | b | o | a |
Remarks: the rhd testing result of above-mentioned 10 samples is feminine gender.
Second group of experiment:
1st, prepare 10 parts of a, b, o and ab blood group peripheral blood, wherein 5 parts is that rhd is negative, in addition 5 parts is that rhd is positive, every part
10ml.
2nd, take above-mentioned peripheral blood 2.5ml to add in the test tube containing coagulant, 300g is centrifuged 2 minutes, take 16 μ l hematocrits red thin
Born of the same parents, add 1984 μ l physiological saline, are configured to 0.8% red cell suspension.
3rd, integrated card and each 50 of conventional card are taken, 200g is centrifuged 2 minutes, tears aluminium film.
4th, it is directly added into 2 μ l peripheral bloods toward in integrated card, add the red cell suspension of 50 μ l simultaneously toward in conventional card.
5th, the conventional card having added red cell suspension is put into 70g in centrifuge to be centrifuged 1 minute, then 100g centrifugation 4 minutes.
6th, the integrated card having added peripheral blood is put into 200g in centrifuge to be centrifuged 2 minutes, then 100g centrifugation 4 minutes.
7th, card taking result of determination, result such as following table two.
Blood group pattern detection result known to table two
From the above experimental results, under the premise of the accuracy ensureing blood type test card, the integral type of the present embodiment
Abo/rhd blood typing detection card can directly add periphery whole blood and be detected, be centrifuged without to periphery whole blood
Take out after red blood cell is diluted and detected again, it is possible to reduce the operating procedure of traditional abo/rhd blood typing detection card,
Improve efficiency.
Above content is to further describe it is impossible to assert with reference to specific preferred embodiment is made for the present invention
Being embodied as of the present invention is confined to these explanations.For those skilled in the art, do not taking off
On the premise of present inventive concept, some equivalent substitutes or obvious modification can also be made, and performance or purposes are identical, all answer
When being considered as belonging to protection scope of the present invention.
Claims (8)
1. a kind of micro-pipe for Blood grouping it is characterised in that: described micro-pipe up comprises gel, antibody work successively from bottom
Make liquid and serum separation gel；Dilute with the antibody making described blood group antibody holding activity containing blood group antibody in described antibody working solution
Release liquid；The proportion of described serum separation gel, between 1.075～1.077, can be sent out under the centrifugal action of 150g～300g
Raw thixotroping, becomes fluid by solid；The proportion of described antibody diluent is more than serum separation gel and is less than red blood cell；Described serum
Separation gel includes each component of following weight portion: cyclopentadiene oligomers 35-45 part, organic gelling agent 12-25 part, organic gelling
Agent dispersant 15-30 part, viscosity reductant 8-12 part and proportion conditioning agent 8-12 part；Described organic gelling agent is dibenzylidene sorbitol
Alcohol；Described organic gelling agent dispersant be polyox-yethylene-polyoxypropylene block copolymer, 1-Methyl-2-Pyrrolidone, dmso,
At least one in dma and dmf；Described viscosity reductant be in phthalic acid two (2- ethylhexyl) ester and epoxy resin at least
A kind of；Described proportion conditioning agent is the silica of 5 μm -10 μm of particle diameter, titanium dioxide, the chlorinated paraffin of chlorination degree 40%, chlorine
The chlorinated paraffin of change degree 70%.
2. micro-pipe as claimed in claim 1 it is characterised in that: the proportion of described antibody diluent between 1.078-1.082,
Ph value is between 6.6-6.8.
3. micro-pipe as claimed in claim 1 it is characterised in that: described gel be sephadex, particle diameter be 40-60 μm.
4. micro-pipe as claimed in claim 1 it is characterised in that: in described micro-pipe, the volume of described antibody working solution and institute
The weight ratio stating gel is between 1:3 to 1:5；The addition of described serum separation gel is 0.8g-2g.
5. a kind of integral type abo/rhd blood typing detection card it is characterised in that: on described detection card, there are 8 micro-pipes, described
8 micro-pipes are respectively:
First micro-pipe: the blood group antibody containing in the micro-pipe described in claim 1-4 any one, and described antibody working solution
For anti-a monoclonal antibody；
Second micro-pipe: the blood group antibody containing in the micro-pipe described in claim 1-4 any one, and described antibody working solution
For anti-b monoclonal antibody；
3rd micro-pipe: the blood group antibody containing in the micro-pipe described in claim 1-4 any one, and described antibody working solution
For anti-d monoclonal antibody；
4th micro-pipe: up comprise gel, antibody diluent and serum separation gel successively from the bottom of described 4th micro-pipe；
5th micro-pipe～the 8th micro-pipe: repeat the first micro-pipe～the 4th micro-pipe respectively.
6. a kind of preparation method of integral type abo/rhd blood typing detection card is it is characterised in that comprise the steps of:
(1) prepare antibody diluent, the proportion of described antibody diluent is more than serum separation gel and is less than red blood cell；
(2) gel swelling: using physiological saline or described antibody diluent, gel is carried out swelling；
(3) prepare antibody working solution: respectively anti-a monoclonal antibody, anti-b monoclonal antibody and anti-d monoclonal antibody are pressed every 1 body
The antibody of long-pending part adds the described antibody diluent proportions of 7 parts by volume to become described antibody working solution；
(4) prepare specific gel: add the ratio of antibody working solution described in 1.2-1.7ml according to swelling gel described in every 1g
Example is configured to respectively have the anti-monoclonal anti-a gel of igm property, has the anti-monoclonal anti-b gel of igm property and have
The anti-monoclonal anti-d gel of igm property；And preparation neutral gum: add 1.2-1.7ml according to swelling gel described in every 1g
The proportions of described antibody diluent become neutral gum；Wherein: anti-a antibody titer >=128, anti-b monoclonal antibody effect
Valency >=128, anti-d antibody titer >=128；
(5) dispense: according to the filling amount of every pipe 45-55 μ l, by anti-for described anti-monoclonal a gel, anti-monoclonal anti-b gel, resist
Monoclonal anti-d gel, neutral gum are added separately in the first micro-pipe, the second micro-pipe, the 3rd micro-pipe and the 4th micro-pipe, the 5th micro-pipe
～the eight micro-pipe repeats the first micro-pipe～the 4th micro-pipe respectively；
(6) prepare serum separation gel, the proportion of described serum separation gel between 1.075～1.077,150g～2000g's
Under centrifugal action, thixotroping can occur, fluid is become by solid；Described serum separation gel includes each component of following weight portion:
Cyclopentadiene oligomers 35-45 part, organic gelling agent 12-25 part, organic gelling agent dispersant 15-30 part, viscosity reductant 8-12 part
With proportion conditioning agent 8-12 part；Described organic gelling agent is dibenzylidene sorbitol；Described organic gelling agent dispersant is poly-
Oxygen ethene-polyoxypropylene block copolymers, 1-Methyl-2-Pyrrolidone, at least one in dmso, dma and dmf；Described subtract
Stick is at least one in phthalic acid two (2- ethylhexyl) ester and epoxy resin；Described proportion conditioning agent is particle diameter 5 μ
M-10 μm of silica, titanium dioxide, the chlorinated paraffin of chlorination degree 40%, the chlorinated paraffin of chlorination degree 70%；
(7) the described serum separation gel of 0.8g-2g, 150g-300g centrifugation one are added respectively in the first micro-pipe～the 8th micro-pipe
Fix time, make described serum separation gel occur thixotroping to be become by solid and catch after fluid on the interface of described antibody working solution；
(8) sealing, labeling and encapsulation.
7. preparation method as claimed in claim 6 is it is characterised in that described step (6) specifically includes following steps:
(61) by 35-45 part cyclopentadiene oligomers heating and melting；
(62) organic gelling agent 12-25 part, organic gelling agent dispersant 15-30 part are added in the cyclopentadiene oligomers melting
With viscosity reductant 8-12 part, kneaded mixture 20-40 minute；
(63) it is cooled to normal temperature, add proportion conditioning agent 8-12 part, vacuum kneading mixing under the vacuum of 0.1bar-0.5bar
Thing 20-40 minute.
8. preparation method as claimed in claim 6 is it is characterised in that the described antibody diluent of preparation in described step (1)
Process as follows: in every 1l pure water add 1.2-1.7g sodium chloride, 12-30g glycine, 0.2-0.3g disodium hydrogen phosphate,
0.46-0.56g sodium dihydrogen phosphate, 10-20g bovine serum albumin(BSA), 12-25g glycerine, 0.45-0.68g methyl p-hydroxybenzoate
With 0.1-0.16g propylparaben.
Priority Applications (3)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510601037.4A CN105223366B (en) | 2015-09-18 | 2015-09-18 | Micropipe for blood grouping, one-piece ABO/RhD blood typing test card for blood grouping and preparation thereof |
| HK16103536.7A HK1215813A1 (en) | 2015-09-18 | 2016-03-25 | Microtube for detecting blood groups, an integrated abo/rhd blood groups detection card and its preparation abo/rhd |
| PCT/CN2016/078941 WO2017045397A1 (en) | 2015-09-18 | 2016-04-11 | Micropipe for blood grouping, one-piece abo/rhd blood typing test card, and preparation therefor |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510601037.4A CN105223366B (en) | 2015-09-18 | 2015-09-18 | Micropipe for blood grouping, one-piece ABO/RhD blood typing test card for blood grouping and preparation thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN105223366A CN105223366A (en) | 2016-01-06 |
| CN105223366B true CN105223366B (en) | 2017-01-18 |
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| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201510601037.4A Active CN105223366B (en) | 2015-09-18 | 2015-09-18 | Micropipe for blood grouping, one-piece ABO/RhD blood typing test card for blood grouping and preparation thereof |
Country Status (3)
| Country | Link |
|---|---|
| CN (1) | CN105223366B (en) |
| HK (1) | HK1215813A1 (en) |
| WO (1) | WO2017045397A1 (en) |
Families Citing this family (11)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105223366B (en) * | 2015-09-18 | 2017-01-18 | 深圳市达科为生物工程有限公司 | Micropipe for blood grouping, one-piece ABO/RhD blood typing test card for blood grouping and preparation thereof |
| CN105586251B (en) * | 2016-03-18 | 2018-02-27 | 深圳市达科为生物工程有限公司 | A kind of separation of lymphocytes pipe and preparation method thereof, separation of lymphocytes method |
| CN108845139A (en) * | 2018-05-04 | 2018-11-20 | 江苏中济万泰生物医药有限公司 | Antibody sample dilution saves liquid and preparation method thereof |
| CN109991408B (en) * | 2019-04-10 | 2022-07-01 | 福州迈新生物技术开发有限公司 | Antibody diluent composition, antibody diluent and preparation method thereof |
| CN111337693A (en) * | 2020-03-30 | 2020-06-26 | 江苏中济万泰生物医药有限公司 | Preparation method of Rh blood type detection card |
| CN112577796B (en) * | 2020-11-02 | 2022-10-21 | 广东省中医院（广州中医药大学第二附属医院、广州中医药大学第二临床医学院、广东省中医药科学院） | Erythrocyte heat diffusion elution kit and application thereof in ABO blood type identification |
| CN112444518B (en) * | 2020-11-18 | 2022-11-22 | 天津德祥生物技术股份有限公司 | Blood type reagent card centrifugal detection device |
| CN112684191A (en) * | 2020-12-04 | 2021-04-20 | 上海润普生物技术有限公司 | ABO blood type positive and negative shaping and Rh blood type detection card and preparation method thereof |
| CN112684190A (en) * | 2020-12-04 | 2021-04-20 | 上海润普生物技术有限公司 | ABO blood type positive typing and RhD blood type detection card and preparation method thereof |
| CN112684176A (en) * | 2020-12-04 | 2021-04-20 | 上海润普生物技术有限公司 | Rh blood group antigen detection card and preparation method thereof |
| CN113189354B (en) * | 2021-04-30 | 2023-09-19 | 重庆国际旅行卫生保健中心(重庆海关口岸门诊部) | Portable quick blood typing detection assembly |
Family Cites Families (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| DE69627627T2 (en) * | 1995-08-28 | 2004-02-05 | Sekisui Kagaku Kogyo K.K. | COMPOSITION FOR SEPARATING SERUM OR PLASMA |
| ATE423312T1 (en) * | 2001-12-04 | 2009-03-15 | Sekisui Chemical Co Ltd | COMPOSITION FOR SEPARATING BLOOD SERUM OR PLASMA AND VESSEL CONTAINING SAME FOR BLOOD EXAMINATION |
| US20070054405A1 (en) * | 2003-10-23 | 2007-03-08 | Ortho-Clinical Diagnostics, Inc. | Patient sample classification based upon low angle light scattering |
| US20120202225A1 (en) * | 2011-02-01 | 2012-08-09 | Arryx, Inc. | Methods and devices for immunodiagnostic applications |
| CN102680716A (en) * | 2012-06-07 | 2012-09-19 | 北京金豪制药股份有限公司 | ABO/RhD blood group antigen detection reagent card and preparation method thereof |
| CN203083997U (en) * | 2012-12-13 | 2013-07-24 | 江苏创生生物技术有限公司 | ABO and RhD blood group antigen detection card |
| CN104569444B (en) * | 2014-12-22 | 2016-03-30 | 合肥天一生物技术研究所 | A kind of positive reverse type of ABO and RhD blood type test card |
| CN105223366B (en) * | 2015-09-18 | 2017-01-18 | 深圳市达科为生物工程有限公司 | Micropipe for blood grouping, one-piece ABO/RhD blood typing test card for blood grouping and preparation thereof |
-
2015
- 2015-09-18 CN CN201510601037.4A patent/CN105223366B/en active Active
-
2016
- 2016-03-25 HK HK16103536.7A patent/HK1215813A1/en unknown
- 2016-04-11 WO PCT/CN2016/078941 patent/WO2017045397A1/en active Application Filing
Also Published As
| Publication number | Publication date |
|---|---|
| WO2017045397A1 (en) | 2017-03-23 |
| HK1215813A1 (en) | 2016-09-15 |
| CN105223366A (en) | 2016-01-06 |
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